产品介绍：
PepMute™ Plus体外siRNA和DNA转染试剂是PepMute™转染试剂(目录号：SL100566)的升级版。 PepMute™ Plus体外siRNA和DNA转染试剂是在PepMute™转染试剂的基础上经过其骨架添加输水性氨基酸所得到的。基于我们独特的pH依赖的分子构象转化技术，我们在36个氨基酸组成的骨架多肽基础上经过添加输水性的氨基酸并经过反复筛选得到，这种化学修饰赋予PepMute™ Plus试剂一种独特性质，即在形成转染复合物时该试剂获得了自我装配的特性。这一特性使得该试剂成为一个最为高效的转染试剂，实验证明，该试剂对siRNA有及其高效的转染效率，即使在siRNA终浓度为1.0 nM时，也可以获得95%基因沉默效率。 PepMute™ Plus试剂已经被证明可有效地对多种哺乳动物细胞进行有效的siRNA和DNA转染以及和DNA/siRNA共转染。
Figure 1. A cartoon showing PepMute™ Plus siRNA Transfection Reagent was developed
Size & content:
– PepMute™ Plus Transfection Reagent, 1.0 ml, sufficient for ~1000 reactions based on transfecting 2.5 pmols siRNA in 24-well plate
– PepMute™ Plus Transfection Buffer(5x ), formulated for maximal transfection efficiency, 8.0 ml
Storage:
Store at 4 °C and never freeze. If stored properly, the product is stable for 12 months or longer.
Advantages:
– Effective knockdown (95%) with only 0.5 pmol siRNA (1.0 nM siRNA in 24-well plate)
– Formulated for hard-to-transfect mammalian cells
– One tube reaction and easy transfection protocol
– Best for broad spectrum especially hard-to-transfect mammalian cells
– Very low cytotoxicity
Comparisons of Knockdown Efficacy of PepMute™ Plus siRNA & DNA Transfection Reagent with Brand Name Products
Figure 2. Knockdown efficacy comparison of PepMute™ Plus Transfection Reagent (upper panel) vs. Dharmafect™ 4 siRNA Transfection Reagent (middle panel) and Lipofectamine™ 2000 (lower panel) on HEK293 cells. siRNA targeting renilla luciferase at different final concentrations ranging from 0.5 to 20 nM was co-transfected with renilla luciferase gene (0.5 µg of pRL-CMV DNA per well) by the above three transfection reagents per manufacturers’ protocols into HEK293 cells growing on a 24-well plate. Renilla luciferase activity was determined 24h after post co-transfection with renilla luciferase determination system (Promega). The luminescence was measured from 5.0 µl of lysate during 10s integration with a luminometer (Beckman Coulter LD 400). Luciferase activity was expressed as light units integrated over 10s (RLU) and normalized per mg of cell protein by using the BCA assay. The errors bars represent standard deviation derived from triplicate experiments. Luciferase-silencing efficiency was calculated relative to untreated cells. While PepMute™ Plus and Dharmafect™ 4 reagents delivered significant gene silencing from 1.0 nM of renilla luciferase siRNA, lipofectamine™ 2000 gave good knockdown only after 30 nM (data not shown).
Figure 3. PepMute™ Plus Transfection Reagent knocked down GFP gene expression in HepG2 cells. GFP cDNA, pEGFP-N3, was co-transfect with a siRNA targeting GFP gene (final 5.0 nM, right panel) and a sham siRNA (final 5.0 nM, left panel) into HepG2 cells by PepMute™ Plus Transfection Reagent. GFP fluorescence was visualized 24h post transfection with a Nikon T2000 fluorescence micorscopy. Quantitative analysis showed that GFP siRNA at 5.0 nM delivered by PepMute™ Plus Transfection Reagent knocked down 96% co-transfected GFP expression in HepG2 cells.
转染操作步骤
– A Protocol for DNA/siRNA Co-Tranafection to Mammalian Cells 
– A Protocol for siRNA Transfection to Mammalian Cells
– A Protocol for DNA Transfection to Mammalian Cells
